Laboratory of Biomolecular Structure and Function

In This Section

Introduction: Three-dimensional structures of proteins determined by X-ray crystallography provide the atomic-scale detail needed to design better therapies to treat diseases and improve human health. After the discovery of the initial crystals for a protein of unknown structure, researchers often must optimize the diffraction quality of the crystals. This step requires testing many different conditions. The human proteins studied at OUHSC often have post-translational modifications that complicate protein preparation and crystallization. The Laboratory of Biomolecular Structure and Function (LBSF) provides a local source of X-ray diffraction instruments with cryosystems that shorten the crystal optimization step. The LBSF also provides help with protein expression, purification, crystallization, data collection with synchrotron radiation, small angle x-ray scattering studies, molecular modeling, and structure-based drug design. In addition, the LBSF interfaces with the BMB Physical Biochemistry Facility for further biophysical characterization of pure protein (ligand binding and thermal stability studies).

Funding sources: The LBSF is supported by a combination of the OUHSC Office of the Vice President of Research, the OUSHC Department of Biochemistry and Molecular Biology, the NIH (via the OCSB and the Ok-INBRE), and user fees. The LBSF is funded in part by grants from the National Institutes of General Medical Sciences of the National Institutes of Health:

National Institute of General Medical Sciences of the National Institutes of Health (Award No. P20GM103640 PI Ann West)

National Institute of General Medical Sciences of the National Institutes of Health (Award No. P20GM103447 PI Darrin Akins)

X-ray diffraction:
The laboratory itself is equipped with modern crystallographic
equipment and computers: Rigaku Micromax 007 X-ray generator, R-AXIS IV and Mar345
image plate detectors, VariMax optics on both ports, Rigaku and Oxford
Systems crystal cooling systems. The R-AXIS IV has an optional helium
path for working with large unit cells.
D-Trek, XDS, Mosflm and HKL2000 image processing software are available for data processing. Phenix, CCP4, or CNS program suites can be used for structure determination and refinement, while COOT, Chimera, Pymol, VMD, and Molmol provide graphical representations of the results in 2D and 3D.

Biophysical studies: A Wyatt
plate-reading dynamic light scattering instrument enables
high-throughput data collection with no user intervention for detection
for the presence of aggregates, for evaluation of the homogeneity of
the sample, and for determining the oligomerization state of the protein in solution.
This instrument is useful for screening samples and buffer conditions
prior to crystallization experiments and other biophysical experiments
that require a homogeneous sample such as small-angle X-ray scattering
(SAXS), microcalorimetery, and microscale thermophoresis. The DLS
instrument was purchased with funds from both the COBRE and the INBRE.
Cite both grants in publications.

Protein production: The LBSF oversees a Protein
Expression and Purification facility where users can overexpress and
purify proteins on the milligram scale for crystallographic or other
kinds of studies. Training in the use of the instruments is provided by
Dr. Terzyan.

Molecular Modeling: Recently a Linux workstation
with 4 GTX1080 GPUs (one GPU is equivalent of 138 CPUs) was purchased by
the OCSB to carry out molecular dynamics simulations of macromolecules
and other computationally intensive molecular modeling work. Available
software includes
AMBER16, Gromacs, and Rosetta software.

Workshop on SAXS data analysis July 28, 2017 at OUHSC (23 people from OU, OUHSC, and OSU attended). Lead by Angela Criswell (Rigaku Inc.) and Blaine Mooers.

Five-week course on practical structure-based drug design offered in the third session of Spring semester 2018.

Art Robbin Crystallization Robot installed in May 2018.

To acknowledge COBRE support if you used any instrument or service of the LBSF:

"Research reported in this [publication, release] was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103640."

To acknowledge the INBRE support if you used the dynamic light scattering instrument:

"Research reported in this [publication, release] was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103447."